Pharmaecutical composition containing silybin, ve and l-carnitine

ABSTRACT

A pharmaceutical composition. The pharmaceutical composition is prepared from the following raw materials in parts by weight: 8.75-60 parts of silybin, 15-65 parts of phospholipid, 25-200 parts of a Pu&#39;er tea extract, 6.25-40 parts of vitamin E, and 8.3-60 parts of L-carnitine. The composition can be used for treating non-alcoholic fatty liver diseases.

TECHNICAL FIELD

The present invention relates to the technical field of medicines, andparticularly to a pharmaceutical composition containing silybin for thetreatment of hepatopathy.

BACKGROUND ART

In the late 1960s and 1980s, the pharmaceutists of West Germany withH.wagner as representative extracted the active ingredient from thefruit of the Silybummarianum, which is named as silymarin, a new classof flavonoid having a C-9 substituents, i.e., a flavonoid lignanscondensed with a dihydroflavonol and a phenylpropanoid derivative.Silybin(silibinin) is one of the main components of silymarin.Pharmacological and toxicological studies have shown that silybin hasthe effects of protecting and stabilizing the hepatocyte membrane,promoting the recovery of hepatocyte and improving the liver function.Silybin has different levels of protection and treatment effects onvarious types of hepatic injury caused by hepatic poisons such as carbontetrachloride, thioacetamide, hydroxycholine, phalloidine, mucronatine,etc. And silybin can be used for treating acute and chronic hepatitis,early hepatocirrhosis, fatty liver, toxic or drug-induced hepatopathy.

The silybin is poor in water solubility and common, organic solvents,resulting in low bioavailability and thereby affecting the clinicalefficacy. To improve the bioavilability thereof, domestic and externalpharmacy workers have made substantial amounts of work. The measures toimprove the absorption of poorly soluble drugs are typically superfinegrinding, salinization, and the addition of cosolvent, etc. In recentyears, the studies have shown that the dissolution and bioavailabilityare greatly improved by the methods of formulating into cyclodextrininclusion compound, solid dispersion, synthetic phospholipid complex andformulating into different dosage forms.

From the perspective of solid preparation, the phospholipid complex is amore specific solid dispersion, which has a fixed melting point, is amolecular compound (complex) whose chemical nature is more stable anddifferent from the compound of drug and phospholipid, such compoundsvaries with the types of phospholipid and ratios of drug tophospholipid, and a phospholipid molecule can be bound with a differentnumber of drug molecules. Deduced from the spectroscopy characteristicsof the complex, the drug has a strong interaction with the polar groupsof the phospholipid, which inhibits the free rotation of the singlechains in the molecule, whereas the two long fatty acid chains of thephospholipid do not participate in the complex reaction and are free toshift and wrap the polar portions of the phospholipid to form alipophilic surface, so that the complex shows strong lipid solubility.The complex changes the physiochemical properties of drug, and thusincreases the lipid solubility of the drugs and reduces the watersolubility of the drugs, and promotes the combination of drug moleculesand cell membranes to improve the absorption and increases thebioavailability of tire drag.

Pu'er tea is a unique and famous tea in Yunnan province. The localityhas moderate climate, abundant rainfall and is mist-shrouded. Pu'er teais divided into two series by Yunan big leaf species sun-dry tea midreprocessing thereof: the unzymic Pu'er tea by directly re-processinginto the finished product and the enzymic Pu'er tea by re-processingafter the artificial accelerated fermentation, and the patterns of whichare divided into loose tea and compressed tea; natural aging process isalso persistently carried out after the finished products, with theunique qualities gets better.

Pu'er tea is the only post-fermented tea, and substances harmful to thehuman body such as theophylline, tea polyphenols are degraded in thelong process of fermentation, so the product is mild, does not stimulatethe body, and also can promote metabolism, accelerate the digestion andtransformation of fats and toxins in the body. For the problems ofobesity and three-hypes which are puzzling urbanites, Pu'er tea can playa good mitigation effect, such as expelling of toxin, nourishing thestomach, anti-inflammatory, reducing the cholesterol, off lipid andremoving grease, cosmetic slimming. Modern technologies show that Pu'ertea can improve insulin resistance, regulate levels of blood lipid andleptin, etc., and cart block the fat accumulation of hepatic parenchymalcell caused by insulin resistance to some extent.

Non-alcoholic fatty liver disease (NAFLD) is a metabolic stress-inducedhepatic injury that is closely related to insulin resistance and geneticsusceptibility, the pathological changes of which are similar toalcoholic fatty liver disease. NAFLD is a clinicopathological syndromecharacterized by steatosis and fat storage of hepatocytes in the hepaticlobule but without history of alcohol abuse. NAFLD shows differentdegrees of hepatic lesion, from simple fatty liver without anyinflammation to severe inflammatory response of severe fibrosis and evencirrhosis, mainly includes 3 types: simple fatty liver, steatohepatitis,fatty cirrhosis.

Non-alcoholic fatty liver disease treatment:

1. Prevention of protopathies or associated risk factors.

2. Basal treatment: developing a reasonable energy intake and dietadjustment, taking moderate aerobic exercises, correcting bad lifestylesand behaviors.

3. Avoiding aggravating hepatic injury: preventing a sharp decline inweight, drug abuse and other factors that may induce exacerbation ofhepatopathy.

4. Weight loss: requiring all NAFLD patients who are overweight, andhave visceral obesity and rapid weight gain is the short term to changethe lifestyles to control weight and reduce waist circumference. Basaltreatment for 6 months, weight loss <0.45 kg per month, or body massindex (BMI) >27 kg/m² combined with blood lipid, blood glucose, bloodpressure and other indicators of more than two abnormalities mayconsider adding sibutramine or orlistat and other obesity drugs, weightloss per week should not exceed 1.2 Kg (children do not exceed 0.5 Kgper week); BMI>40 kg/m² or BMI>35 kg/m² combined with sleep apneasyndrome and other obesity-related diseases, may consider the proximalend gastric bypass procedures to lose weight (II-1, II-2, II-3, III).

5. Insulin sensitizer: combined with type 2 diabetes, impaired glucosetolerance, fasting plasma glucose and visceral obesity, may consider theapplication of metformin and thiazolidinediones in order to improveinsulin resistance and control of blood glucose (II-1, II-2, II-3),

6. Hypolipidemic agents: dyslipidemia, with basal treatment and (or)application of weight loss and hypoglycemic drugs for more than 3-6months, is still mixed with hyperlipidemia or hyperlipidemia, combinedwith more than 2 risk factors, should consider adding the use offibrates, statins or protocol and other hypolipidemic drugs (II-1, II-2,II-3).

7. Drags for hepatopathy; NAFLD associated with hepatic dysfunction,metabolic syndrome, 3-6 months after basal treatment remainsineffective, and liver biopsy shows NASH and chronic progression of thecourse of the disease, the drug auxiliary treatment for hepatopathy canbe used with antioxidant, anti-inflammatory, anti-fibrosis, and relateddrugs (II-1, II-2, II-3, III) such as polyene phosphatidylcholine,vitamin E, silymarin and ursodeoxycholic acid can be rationally chosenaccording to drug performance, disease activity and stage of thedisease, but multi-drags should not be applied simultaneously.

8. Liver transplantation: mainly for NASH-related end-stage hepatopathyand some cryptogenic J hepatocirrhosis, and the metabolic condition(III) should be screened before liver transplantation. BMI>40 kg/m² iscontraindication to liver transplantation (III).

The above treatments have not been used by being mixed together, such asa combination of hypoglycemic and hepatopathy drugs, or a combination oflipid-lowering and hepatopathy drugs. Therefore, the search for apharmaceutical with a variety of health-promoting functions cannot wait.

SUMMARY OF THE INVENTION

In order to solve the above technical problems, the present inventionprovides a pharmaceutical composition and a preparation thereof, whichis therapeutically effective for non-alcoholic fatty liver diseases.

The present invention provides the methods for preparing thepharmaceutical composition and the preparation thereof.

The present invention is achieved by the following technical solutions:

A pharmaceutical composition is prepared from the following bulk drugsby weight ratio:

-   -   8.75-60 parts of silybin    -   15-65 parts of phospholipid    -   25-200 parts of Pu'er tea extract    -   6.25-40 parts of vitamin E    -   8.3-60 parts of L-carnitine.

It is preferably prepared from the following bulk drugs by weight ratio:

-   -   25-40 parts of silybin    -   30-50 parts of phospholipid    -   80-120 parts of Pu'er tea extract    -   0-30 parts of vitamin E    -   25-40 parts of L-carnitine.

It is most preferably prepared from the following bulk drags by weightratio:

-   -   35 parts of silybin    -   42 parts of phospholipid    -   100 parts of Pu'er tea extract    -   16.7 parts of vitamin E    -   33.3 parts of L-carnitine.

The phospholipid of the present invention is a phospholipid or lecithin,preferably soybean phospholipid, which is mainly composed ofphosphatidylcholine.

The rote of the phospholipid in the present invention is to promote thedissolution and absorption of medicaments, silybin is a medicament withlow solubility and low permeability, and the phospholipid is combinedthere with to form a phospholipid complex so as to improve solubility ofthe silybin, thereby improving the bioavailability of the drugs.

Described silybin and phospholipid are both known from the prior art orcommercially available. In order to better exert the efficacy of thepresent invention, the silybin of the present invention is preferablyprepared by dissolving silymarin in 80% ethanol, filtering and washingthe precipitate with 95% ethanol for three times, collecting Sheprecipitate. The precipitate is dissolved in anhydrous ethanol,filtered, and the filtrate is added with a certain amount of water toseparate out the precipitate, and the precipitate is collected byfiltration, dried under reduced pressure, pulverized and mixed.

The Pu'er tea extract is commercially available, preferably a DEEPURE®Pu'er tea essence. It is also possible to be prepared according to theprior art. In order to better exert the efficacy of the presentinvention, the Pu'er tea essence or Pu'er tea extract is preferablyprepared according to the method of patents (publication No.CN101961061A, CN101961061B, CN101961425A, CN101961425B, CN101961060A,CN101961059A, CN101961059B).

For example, said Pu'er tea essence is prepared as follows:

Step 1, Pu'er tea leaves are decocted with 6-12 times the volume ofwater for 2-4 times, 0.5-2hours each time; extract solution is filtered,and filtrate is concentrated under reduced pressure and the temperatureof ≦70° C. to tire weight of tea leaves:the volume ofconcentrate=1:2-1:3;

Step 2, the concentrate is centrifuged with a centrifuge, thecentrifugate is concentrated under reduced pressure to density of1.1-1.25 at 45-65° C., concentrated cream is spray dried or microwavedried to obtain the final product.

Preferably, the steps are present as follows:

Step 1, Pu'er tea leaves are decocted with 6-12 times the volume ofvigorously boiling water for 3 times, 0.5-2 hours each time; extractsolution is filtered, and filtrate is concentrated under reducedpressure and the temperature of ≦70° C. to the weight of tea leaves:thevolume of concentrate=1:2-1:3;

Step 2, the concentrate is centrifuged with a tripod pendulum type batchcentrifugal, the tripod pendulum is centrifuged with a tubular-bowlcentrifuge, and the centrifugate is concentrated under reduced pressureto density of 1.1-1.25 at 45-65° C., concentrated cream is spray driedor microwave dried to obtain the final product;

wherein tubular-bowl centrifuge condition is; centrifuge speed:15000-19000 rpm/min; spray drying conditions are: inlet temperature:140-190° C., outlet temperature: 75-95° C.

Most preferably, the steps are present as follows:

Pu'er tea leaves are decocted with vigorously boiling water for 3 times,the first time decocted for 1.5 h, 10 times the volume of water added;the second time decocted for 1.5 h, 8 times the volume of water added;the third time decocted for 1 h, 8 times the volume of water added,extract solution is filtered, and filtrate is concentrated under reducedpressure and the temperature of ≦70° C. to the weight of tea leaves:thevolume of concentrate=1:2-1:3, the concentrate is centrifuged with atripod pendulum type batch centrifugal, the tripod pendulum iscentrifuged with a tubular-bowl centrifuge, and the centrifugate isconcentrated under reduced pressure to density of 1.1-1.25 at 45-65° C,concentrated cream is spray dried or microwave dried to obtain the finalproduct; wherein tubular-bowl centrifuge condition is: centrifuge speed:15000-19000 rpm/min; spray drying conditions are: inlet temperature:140-190° C., outlet temperature: 75-95° C.

The vitamin E is any one of commercially available vitamin E for dietarysupplementation or drug use, vitamin E acetate and vitamin E succinate.

The L-carnitine is either commercially available L-carnitine for dietarysupplement or pharmaceutical use or L-carnitine tartrate.

The above compositions are made by weight ratios, and may be increasedor reduced according to corresponding proportion in productionprocesses, such as large-scale production can be in unit of kg or T(ton); small scale preparations can also be in unit of g. The weight canbe increased or reduced, but the proportions of the weight ratio of bulkdrugs between the components remain unchanged.

The proportions of the above weight ratio are obtained throughscientific screening, for special patients, such as patients with severeor mild symptom, obese or thin patients, the proportions of the amountof composition can be accordingly adjusted, increased or decreased nomore than 10%, the efficacy is substantially constant.

Any pharmaceutically acceptable dosage forms can be formulated in theformulation of a pharmaceutical preparation, the dosage forms areselected from: tablet, sugar coated tablet, film coated tablet, entericcoated tablet, capsule, hard capsule, soft capsule, oral liquid, oralagent, granule, pill powder, paste, sublimed preparation, suspensionagent, solution, injection, suppository, ointment, emplastrum, creme,spray, patch. Preferably oral preparations, and optimal preferablytablet, capsule, granule.

Some pharmaceutically acceptable carriers can be added into thecompositions of the present invention as needed, the pharmaceuticalpreparations can be prepared using galenic pharmacy conventionaltechniques, such as mixing the pharmaceutically active substances withpharmaceutically acceptable carriers. The pharmaceutically acceptablecarriers are selected from: mannitol, sorbitol, sorbic acid or sylvite,sodium metabisulfite, sodium bisulfite, sodium thiosulfate, cysteinehydrochloride, mercaptoacetic acid, methionine, vitamin A, vitamin C,vitamin E, vitamin D, azone, disodium EDTA, calcium disodium EDTA, thecarbonate, acetate, phosphate of monovalence alkali metal or aqueoussolution thereof, hydrochloric acid, acetic acid, sulfuric acid,phosphoric acid, amino acid, sodium chloride, potassium chloride, sodiumlactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch,sucrose, lactose, mannitol, silicon derivative, cellulose and derivatethereof, alginate, gelatin, polyvinyl pyrrolidone, glycerine, propyleneglycol, ethanol, Tween 60-80, Span-80, beeswax, lanolin, liquidparaffin, cetyl alcohol, gallic acid esters, agar, triethanolamine,basic amino acid, urea, allantoin, calcium carbonate, calciumbicarbonate, surfactant, polyethylene glycol, cyclodextrin,beta-cyclodextrin, phospholipid material, kaolin, talc, calciumstearate, magnesium stearate, etc. Preferably, the carrier is one ormore of microcrystalline cellulose, lactose, starch, sodiumcarboxymethylcellulose, low substituted hydroxypropyl cellulose, talc.

When the composition of the present invention is prepared intomedicament, the unit dosage of the medicament may contain 0.1-1,000 mgof the pharmaceutically active substance of the present invention, andthe remainders are pharmaceutically acceptable carriers. Thepharmaceutically acceptable carriers may be 0.1-99.9% of the totalpreparation weight by weight. Preferably, the pharmaceuticallyacceptable carriers may be 40-70% of the total preparation weight byweight.

The usage and dosage of the Chinese traditional medicine compositions orpreparations of the present invention are determined according to theconditions of patients while being used.

The cumulative dissolution rate of the pharmaceutical compositionpreparations according to the present invention, such as tablets,capsules, granules and so on, is not less than 60% while dissolution invitro for 2 h and the dissolution rate is greater than or equal to 15%while dissolution in vitro for 30 min, in a dissolution condition:slurry method, rotation speed of 100 rpm and temperature of 37□, andrelease medium is: 1,000 ml of hydrochloric acid solution at pH1.2,dosage: 1 capsule/1 tablet/1 bag granules.

A preparation method of the pharmaceutical composition of the presentinvention comprises the following steps:

{circle around (1)} taking a prescription amount of raw materials forlater use;

{circle around (2)} preparation of silybin complex liquid: weighing aprescription amount of silybin and phospholipid, and dissolving them inthe anhydrous ethanol, heating and refluxing to clarify the solution andcontinuing to heat for a certain time, then concentrating the clearsolution under reduced pressure to a certain volume, to obtain thesilybin complex liquid for later use;

{circle around (3)} granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silybin complex liquidprepared in step {circle around (2)} as a feed liquid, and preparinggranules by a fluidization spray method with a fluidized bed, and dryingafter the liquid complex is all sprayed for later use;

{circle around (4)} total blending: mixing a prescription amount ofvitamin E with the L-carnitine uniformly, and then mixing a mixture ofthe two with the granules prepared in step {circle around (3)} uniformlyin an equal incremental manner to obtain the pharmaceutical composition;

The present invention also includes a preparation step {circle around(5)}, taking the pharmaceutical composition of step {circle around (4)}and pharmaceutically acceptable carriers, and preparing pharmaceuticallyacceptable dosage forms according to the conventional preparationprocess.

Further preferably, the preparation method of the pharmaceuticalcomposition of the present invention, comprises the following steps:

{circle around (1)} taking a prescription amount of raw materials forlater use;

{circle around (2)} preparation of silybin complex liquid: weighing aprescription amount of silybin and phospholipid, and dissolving them inthe anhydrous ethanol, heating and refluxing to clarify the solution andcontinuing to heat for a certain time, then concentrating the clearsolution under reduced pressure to a certain volume, to obtain thesilybin complex liquid for later use;

{circle around (3)} granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silybin complex liquidprepared in step {circle around (2)} as a feed liquid, and preparinggranules by a fluidization spray method with a fluidized bed, and dryingafter the liquid complex is all sprayed in for later use;

{circle around (4)} total blending: mixing a prescription amount ofvitamin E with the L-carnitine uniformly, and then mixing a mixture ofthe two with the granules prepared in step {circle around (3)} uniformlyin an equal incremental manner to obtain the pharmaceutical composition;

{circle around (5)} preparation: taking the pharmaceutical compositionand pharmaceutically acceptable carriers to prepare pharmaceuticallyacceptable dosage forms according to the conventional preparationprocess.

Wherein the heating time described in step {circle around (2)} is0.5-1.5 hours; the concentrated volume is 5%-20% of the original volumeand the temperature of concentration under reduced pressure is 60-80° C.

Wherein the parameters of the fluidized bed in step {circle around (3)}are: the temperature of the materials is 40-65° C., during thegranulation process, the parameters such as fan frequency, inlet airtemperature and infusion frequency are adjusted to keep the materials ingood fluidization state. After completing the granulation, the granulesare dried for 10-60 minutes, and the drying temperature is 55-65° C.

Pu'er tea can improve insulin resistance, regulate the levels of bloodlipid and leptin and other effects, can block the fat accumulation ofhepatic parenchymal cell caused by insulin resistance to a certainextent, combined with the strong free radical scavenging andanti-oxidative stress ability of silybin, the two has preferableanti-NAFLD (non-alcohol fatty liver) effect.

vitamin E is added to further enhance the products' function inbeautifying skin, L-carnitine is added to strengthen the products'function in reducing fat and losing weight.

EXPERIMENTAL EXAMPLE 1 Dissolution Experiment In Vitro

The dissolutions of the silybin-phospholipid-Pu'er tea-VE-L-carnitinecomposition prepared in embodiments 16-20 are determined under thefollowing conditions: the selection of the dissolution methods is basedon the properties of the main component silybin in the compositions, thesilybin, as a medicament with low solubility and low permeability, isthe fourth category in the Biopharmaceutics Classification System (BCS),dissolution and absorption thereof are both the rate-limiting steps,either of which should be resolved at the same time in order to improvethe bioavailability of the drugs. Dissolution stage of the silybin ismainly carried out in the stomach, the absorption stage is mainly earnedout in the small intestine, and measuring the in-vitro dissolution ofthe drug helps to improve fee bioavailability of the drugs. Therefore,the following dissolution method is chosen to evaluate the composition:slurry method, rotation speed of 100 rpm and temperature of 37° C., andrelease medium is: 1,000 ml of hydrochloric acid solution, at pH1.2,dosage: 1 capsule/1 tablet/1 bag granules. The sampling points are: 15,30, 45, 60, 90, 120 min. The cumulative dissolution is determined. Theresults are shown in Table 1 below.

TABLE 1 Accumulated dissolution (%) Time Embodiment EmbodimentEmbodiment Embodiment Embodiment Embodiment Embodiment Embodiment (min)Embodiment 1 18 19 20 21 22 23 25 26 15 19.76% 8.21% 13.78% 15.06% 7.40%8.21% 7.47% 7.09% 9.34% 30 58.78% 21.98% 42.39% 55.75% 28.29% 21.98%28.29% 20.12% 24.56% 45 67.24% 49.44% 68.45% 66.24% 44.33% 49.44% 55.18%52.87% 47.68% 60 79.54% 76.88% 74.88% 69.51% 81.34% 76.88% 74.23% 77.18%74.18% 90 83.97% 79.42% 79.27% 80.96% 82.74% 79.42% 81.32% 80.42% 82.56%120 85.59% 81.83% 80.31% 81.92% 83.14% 81.83% 82.09% 82.08% 84.29%

The dissolution determination of the reference preparation(silybin-phospholipid complex preparation, laboratory homemade) iscarried out, and compared with the silybin-phospholipid-Pu'ertea-VE-L-carnitine compositions prepared in embodiments 19-23, and theresults are as shown, in FIG. 1. It can be seen from the data in Table 1and the graph of FIG. 1:

The in-vitro release of the silybin-phospholipid-Pu'ertea-VE-L-carnitine composition prepared by the preparation method of thepresent invention is significantly better than that of the referencepreparation silybin-phospholipid complex, surprisingly, the cumulativedissolution of the composition in the hydrochloric acid solution atpH1.2 for 2 h reaches to 80%, nearly completely dissolved, which isdoubled as compared with that of the reference preparation, resolvingthe problems of low solubility and low bioavailability of silybin thathave always existed, which will provide the basis for the studies ofdose setting and in-vivo safety and efficacy of silybin composition inthe future.

Pooling the data of in-vitro dissolution experiments and in-vivopharmacological researches, the present invention further improves thedissolution of the pharmaceutica by combining the silybin-phospholipidcomplex with Pu'er tea, vitamin E, and L-carnitine, by continuing theabsorption improvements of the pharmaceutical by increasing thecompatibility of the pharmaceutical and the biofilm after thecombination of silybin and phospholipid, from two aspects of improvingthe dissolution and absorption to improve the bioavailability of themain component silybin.

EXPERIMENTAL EXAMPLE 2 Pharmaceutical Efficacy Test In Vivo

1 Experimental Animals

80 mice with SPF grade and 6-week-old male C57 BL/6J leptin-deficient(ob/ob), 10 mice with SPF grade and 6 weeks old male C57 BL/6J (ob/m),provided by the Beijing Huafukang Bioscience Co., Inc., raised in TaslyInstitute's pharmacological toxicology research center barrier animalroom, at the temperature of 20° C.-25° C., relative humidity of 60%, 5mice in each cage, lighting time of 12 hours, timely and quantitativefeed, ob/ob mice are fed with high fat diet (HFD, D12492), C57 BL/6Jmice are fed with normal diet, both are provided by Beijing HuafukangBioscience Co., Inc., and free chinking water, daily replacement ofpadding.

2 Tested Material

Silybin-phospholipid complex, provided by Tasly Pharmaceutical GroupCo., Ltd., lot number 500902031; Pu'er essence, brown powder, suppliedby Tasly Pharmaceutical Group Co., Ltd., lot number Z001 PE(2014)C06(H);L-carnitine tartrate (containing 68.4% of L-carnitine), provided by theNortheast Pharmaceutical Group Co., Ltd., lot number 0661504001;water-soluble vitamin E, produced by Sigma Company; the above testedsubstances are stored in the sample cabinet of the test room ofPharmacology Institute to be protected from light at room temperature.

3 Experimental Methods

3.1 Experimental Dose Design and Grouping

The silybin-phospholipin complex is administered to the experimentalanimals at a daily dose of 3 g (containing silybin 420 mg, soybeanphospholipid 504 mg); the Pu'er tea extract is administered to theexperimental animals at a daily dose of 1.2 g; The compatibilityproportions and the experimental dose designs of the five differentcompositions are shown in Table 2, the dose of the experimental animalsis set to the corresponding equivalent dose of the corresponding testedmaterials, the formula for calculation is as follows:

animal experimental dose=recommended human dose/60 kg*12.3

TABLE 2 Dose of different compositions Recommended human daily dose (mg)Experimental animal dose design (mg/kg) Formulated Combi- Combi-Combination Combination Combination Combination Combination CombinationCombination Combination medicament nation 1 nation 2 3 4 5 1 2 3 4 5Silybin 420 420 420 105 105 86.10 86.10 86.10 21.53 21.53 Phospholipid504 780 780 195 195 103.32 160.00 160.00 40.00 40.00 Pu'er tea 1200 12002400 300 1200 246.00 246.00 492.00 61.50 246.00 extract Vitamin E 300300 75 150 150 61.50 61.50 15.38 30.75 30.75 L-carnitine 400 400 100 200200 82.00 82.00 20.50 41.00 41.00

3.2 Administration of Tested Substances

After adaptive feeding for 1 week, 80 ob/ob mice of 6-week-old arerandomly divided into 8 groups: a model group, a silybin-phospholipidcomplex group, a Pu'er tea extract group, a combination 1 group, acombination 2 group, a combination 3 group, a combination 4 group, and acombination 5 group, 10 mice for each group. Another 10 6-week-oldC57BL/6J mice are in a normal group. Normal group mice are fed withnormal feed, the model group and the administration group are fed withhigh fat feed (HFD, D12492). In addition, the mice in differentmedicament intervention groups are given the corresponding dose ofdrugs, the doses of the five compositions are as shown in Table 1, thenormal group and the model group are given the same amount of distilledwater, continuous intragastric administration for 6 weeks.

The mice are free to eat and drink during the experiment weekly weight,and the doses are adjusted according to the body weight. After the lastadministration, fasting for 12 h, but water is given, weighing the bodyweight, extracting rats' eyeballs to collect blood and then put to deathby breaking their necks, and the liver is harvested rapidly,physiological saline rinsing, filter paper blotting and preserved in a−20° C. refrigerator after weighing.

3.3 Detecting Indicators and Methods

3.3.1 General Observation

The weights of mice in each group are measured weekly during theexperiment.

3.3.2 Calculation of the Liver Index and Observation of the GeneralMorphology of the Liver

After finishing the experiment, the liver is weighed and the liver indexis calculated, the liver index (%) is liver wet weight/body weight*100%.

3.3.3 Determination of Serum Biochemical Indexes

Blood of all the mice are collected by extracting rats' eyeballs andcentrifuged at 3000 r/min for 15 minutes, the serum is separated andcollected in an EP tube and stored at −20° C. refrigerator for lateruse. The content of glutamic oxaloacetic transaminase (AST),glutamicpyravic transaminase (ALT), total cholesterol (TC), low-densitylipoprotein cholesterol (LDL-C) in serum are measured by 7020 automaticbiochemistry instrument.

3.3.4 Insulin Resistant Index

Serum FINS is detected using the Elisa kit and the insulin resistanceindex is calculated by the formula.

${{Home}\text{-}{IR}} = \frac{{FBG} \times {FINS}}{22.5}$

3.3.5 Liver Histopathological Examination

Frozen sections are prepared from frozen liver tissues and the degree ofhepatic steatosis is observed by oil red O staining. Oil red O stainingoperation steps: frozen slicing→sufficiently washing with distilledwater→staining with oil red O diluent in the dark for 10-15minutes→taking out 6 ml of oil red O saturated liquid, adding 4 ml ofdistilled water, leaving it for 5-10 minutes and filtrating for lateruse→differentiating to interstitial clear under mirror with 60%ethanol→washing with water→nuclear counter staining withhematoxylin→washing with water→sealing piece with neutral gum→microscopeobservation. 3.4 Data Processing

SPSS 15.0 statistical software is used for analysis, the data areexpressed as mean±standard deviation, the t test is used to analyzewhether there's any difference between the two groups before and aftertreatment or not, and the difference is statistically significant withP<0.05.

4 Experimental Results

4.1 The Effects of Each Tested Materials on Body Weight

Mice in each group are measured once a week during the experiment, andthe effects of each group of medicaments on body weight of micesuffering front non-alcoholic fatty liver diseases in each group areobserved. As shown in Table 3, the weight of mice in normal group isincreased slowly and the weight of mice in model group is increased morerapidly. After 6 weeks of administration, except for thesilybin-phospholipid complex group, the other administration groupscould inhibit the weight increases of mice in different degrees(P<0.05), a combined use of silybin-phospholipid complex, Pu'er teaextract, L-carnitine and vitamin E is significantly superior to usingsilybin-phospholipid complex alone.

TABLE 3 Effects of each tested substance on body weight of mice (g, n =10, x ± S) Body weight (g) Before Administration for Administration forAdministration for Group administration 2 weeks 4 weeks 6 weeks Normal24.16 ± 1.46 26.24 ± 2.24 27.47 ± 1.50 27.81 ± 2.03 Model 49.43 ± 3.0355.62 ± 1.95 59.90 ± 2.51 62.41 ± 3.02 Silybin-phospholipid 48.28 ± 2.8855.03 ± 4.08 59.36 ± 5.13 62.48 ± 2.05 complex Combination 1 48.61 ±2.07 50.87 ± 2.82** 52.12 ± 2.56** 54.17 ± 2.29** Combination 2 48.23 ±2.66 52.28 ± 3.10** 55.98 ± 4.54* 58.84 ± 3.52* Combination 3 48.08 ±2.71 51.15 ± 2.82** 53.99 ± 2.15** 56.20 ± 2.47** Combination 4 48.70 ±2.15 50.07 ± 1.85** 50.92 ± 2.37** 52.97 ± 1.82** Combination 5 49.06 ±1.66 51.27 ± 2.66** 52.49 ± 1.77** 54.67 ± 1.89** *compared with themodel group P < 0.05; **compared with the model group P < 0.01;

4.2 Effects of Each Tested Substance on Liver Index

As shown in Table 4, the body weight, liver wet weight and liver indexof the mice in the model group are significantly increased (P<0.01) ascompared with the normal group, and the silybin-phospholipid complexcould decrease the wet weight of the liver (P<0.05), but had nosignificant effect on body weight and liver index of the mice (P>0.05);5 kinds of compositions could reduce the wet weight and liver index ofthe liver of mice (P<0.05), and the effect of a combined use of the fouris significantly better than that of using the silybin-phospholipidcomplex alone.

TABLE 4 Effects of each tested substance on liver index of mice GroupBody weight (g) Liver wet weight (g) Liver index % Normal 27.71 ± 1.941.03 ± 0.11 3.73 ± 0.30 Model 62.23 ± 2.77 4.22 ± 0.34 6.79 ± 0.59Silybin- 62.30 ± 4.79 3.88 ± 0.39* 6.28 ± 0.96 phospholipid complexCombination 1 53.97 ± 2.07** 2.54 ± 0.30** 4.70 ± 0.54** Combination 258.64 ± 3.20* 2.96 ± 0.40** 5.07 ± 0.79** Combination 3 56.02 ± 2.28**2.73 ± 0.43** 4.88 ± 0.88** Combination 4 52.07 ± 1.90** 2.31 ± 0.21**4.44 ± 0.54** Combination 5 54.67 ± 1.90** 2.65 ± 0.25** 4.84 ± 0.47***compared with the model group P < 0.05; **compared with the model groupP < 0.01;

4.3 Effects of Each Tested Substance on Blood Lipid, Liver Function andInsulin Resistance Index

As shown in table 5, the serum TC, LDL, ALT and AST and insulinresistance indexes are significantly increased in non-alcoholic fattyliver model mice compared with the normal group (P<0.05); there is nosignificant improvement in the abnormal elevation of thesilybin-phospholipid complex (P>0.05); the silybin-phospholipid complex,Pu'er tea extract, vitamin E and L-carnitine compatibility group ofdifferent proportions, except for no significant improvement in serumAST by the composition 4 (P>0.05), the other compositions couldsignificantly reduce TC, LDL-C, ALT, AST and insulin resistance indexes(P<0.05), and the effect is better than that of using thesilybin-phospholipid complex and the Pu'er tea extract alone.

TABLE 5 Effects of each tested substance on blood lipid, liver functionand insulin resistance indexes of mice Insulin resistance Group TC LDL-CALT AST indexes Normal  3.03 ± 0.27 0.28 ± 0.05  60.89 ± 34.55 127.78 ±50.91 0.577 ± 0.117 Model 10.27 ± 1.20 2.21 ± 0.54 411.67 ± 95.45 200.89± 26.13 1.172 ± 0.228 Silybin-phospholipid 10.29 ± 1.16 2.26 ± 0.32356.24 ± 17.67 176.49 ± 23.30 1.008 ± 0.385 complex Combination 1  6.86± 1.05** 1.07 ± 0.14** 229.51 ± 90.05** 118.41 ± 20.01** 0.661 ± 0.081**Combination 2  9.12 ± 0.88* 1.71 ± 0.28* 179.51 ± 44.62** 171.74 ±34.80* 0.841 ± 0.262** Combination 3  7.85 ± 1.09** 1.36 ± 0.17** 291.69± 110.33* 158.66 ± 35.36* 0.773 ± 0.135** Combination 4  5.76 ± 1.29**0.97 ± 0.17** 326.03 ± 85.10 110.63 ± 27.18** 0.625 ± 0.089**Combination 5  7.06 ± 1.04** 1.10 ± 0.10** 239.51 ± 86.62** 123.97 ±22.06** 0.753 ± 0.207** *compared with the model group P < 0.05;**compared with the model group P < 0.01;

4.4 Effects of Each Tested Substance on Liver Pathology in Mice

Oil red O staining: according to the size and number of red particles inhepatocytes of liver frozen issues stained by Oil red O under lightmicroscope, it is divided into mild, moderate and severe type. Mild,that is, ⅓-⅔ of red granules are shown per unit area under lightmicroscope, graded as 1 point; moderate, that is, more than ⅔ of thehepatocytes containing red particles, graded as 2 points; severe, thatis, almost all of the hepatocytes containing red particles, graded as 3points; no steatosis is observed, graded as 0 points.

As shown in Table 6, steatosis occurred in nearly all the hepatocytes inthe liver tissues of the model group, and the pathological scores aresignificantly increased than that in the normal group (P<0.01); there isno significant improvement on liver pathological scores by usingsilybin-phospholipid complex or Pu'er tea extract alone (P>0.05); thecombination of different proportions of silybin-phospholipid complex,Pu'er tea extract, vitamin E and L-carnitine can significantly improvethe liver steatosis, reduce the pathological scores (P<0.05), and theeffect is better than that of using the silybin-phospholipid complex andthe Pu'er tea extract alone.

TABLE 6 Effects of each tested substance on liver pathology of miceGroup Oil red O staining pathological score Normal 0.000 ± 0.000 Model2.900 ± 0.316 Silybin-phospholipid complex 2.556 ± 0.726 Pu'er teaextract 2.444 ± 0.726 Combination 1 2.222 ± 0.667* Combination 2 2.400 ±0.516* Combination 3 2.222 ± 0.833* Combination 4 1.778 ± 0.833**Combination 5 2.100 ± 0.738** *Compared with the model group P < 0.05;*Compared with the model group P < 0.01;

5 Experiment Conclusions

The above experimental results show that: the body weight, liver index,blood lipid, ALT, AST and insulin resistance index are significantlyincreased in the mice of the non-alcoholic fatty liver model groupcompared with those in the blank group, and the liver tissues are severesteatosis. Pu'er tea can improve insulin resistance and regulate bloodlipids, L-carnitine also has a better hypolipidemic effect, togetherwith silybin and vitamin E having strong free radical scavenging abilityand antioxidant stress ability, the combination use of the four has asignificant improvement in liver steatosis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an in-vitro release curve, wherein, each sample is: referencepreparation of Shui Lin Jia, Shui Lin Jia without Pu'er tea, andsilybin-phospholipin-Pu'er tea compositions prepared in embodiments16-20.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is further illustrated by the following specificexamples, but is not intended to be limiting of the present invention.

Embodiment 1

Taking 26.25 g of silybin, 45 g of soybean phospholipid, 75 g of Pu'ertea extract, 18.75 g of vitamin E and 25 g of L-carnitine.

{circle around (1)} Preparation of silybin complex liquid: weighing aprescription amount of silybin, soybean phospholipin, and dissolvingthem in the anhydrous ethanol, heating and refluxing to clarify thesolution and continuing to heat for 1 h, then concentrated under reducedpressure and recycling the ethanol to 15% of the original volume forlater use;

{circle around (2)} granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silybin complex liquidprepared in step {circle around (1)} as a feed liquid, preparing thegranules by a fluidization spray method with a fluidized bed,controlling the temperature of materials at 40° C., drying at 60° C. for20 min after the liquid complexes are all sprayed in for later use;

{circle around (3)} mixing a prescription amount of vitamin E with theL-carnitine uniformly, then mixing the mixture of the two with thegranules prepared in step {circle around (2)} uniformly in an equalincremental manner, bagging, made into 1,000 bags of granules.

Embodiment 2

Taking 180 g of silybin, 195 g of soybean phospholipid, 600 g of Pu'ertea extract, 120 g of vitamin E and 180 g of L-carnitine.

{circle around (1)} Preparation of silybin complex liquid: weighing aprescription amount of silybin, soybean phospholipid and dissolving themin the anhydrous ethanol, heating and refluxing to clarify the solutionand continuing to heat for 1.5 h, then concentrated under reducedpressure and recycling the ethanol to 20% of the original volume forlater use;

{circle around (2)} granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silybin complex liquidprepared in step {circle around (3)} as a feed liquid, preparing thegranules by a fluidization spray method with a fluidized bed,controlling the temperature of materials at 65° C., drying at 65° C. for60 min after the liquid complexes are all sprayed in for later use;

{circle around (3)} mixing a prescription amount of vitamin E with theL-carnitine uniformly, and mixing the mixture of the two with thegranules prepared in step {circle around (2)} uniformly in an equalincremental manner, bagging, made into 1,000 bags of granules.

Embodiment 3

Taking 26.25 g of silybin, 195 g of phospholipid, 600 g of Pu'er teaextract, 18.75 g of vitamin E and 25 g of L-carnitine.

{circle around (1)} Preparation of silybin complex liquid: weighing aprescription amount of silybin, soybean phospholipin, and dissolvingthem in the anhydrous ethanol, heating and refluxing to clarify thesolution and continuing to heat for 0.5 hours, then concentrated underreduced pressure and recycling the ethanol to 5% of the original volumefor later use;

{circle around (2)} granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking a silybin complex liquid preparedin step {circle around (1)} as a feed liquid, preparing the granules bya fluidization spray method with a fluidized bed, controlling thematerial temperature at 50° C., drying at 55° C. for 10 min after theliquid complexes are all sprayed in for later use;

{circle around (3)} mixing a prescription amount of vitamin E with theL-carnitine uniformly, and mixing the mixture of the two with thegranules prepared in step {circle around (2)} uniformly in an equalincremental maimer, bagging, made into 1,000 bags of granules.

Embodiment 4

Taking 26.25 g of silybin, 195 g of phospholipid, 75 g of Pu'er teaextract, 12.0 g of vitamin E and 180 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 5

Taking 180 g of silybin, 45 g of phospholipid, 75 g of Pu'er teaextract, 18.75 g of vitamin E and 180 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 6

Taking 180 g of silybin, 45 g of phospholipid, 600 g of Pu'er teaextract, 120 g of vitamin E and 100 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 7

Taking 180 g of silybin, 195 g of phospholipid, 75 g of Pu'er teaextract, 18.75 g of vitamin E and 180 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 8

Taking 26.25 g of silybin, 48.75 g of phospholipid, 75 g of Pu'er teaextract, 37.5 g of vitamin E and 50 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 9

Taking 26.25 g of silybin, 48.75 g of phospholipid, 300 g of Pu'er teaextract, 37.5 g of vitamin E and 50 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 10

Taking 52.5 g of silybin, 97.5 g of phospholipid, 300 g of Pu'er teaextract, 9.375 g of vitamin E and 12.5 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 11

Taking 75 g of silybin, 90 g of phospholipid, 240 g of Pu'er teaextract, 30 g of vitamin E and 75 g of L-carnitine, and preparing 1,000packages of granules according to the method of Embodiment 1.

Embodiment 12

Taking 90 g of silybin, 108 g of phospholipid, 270 g of Pu'er teaextract, 60 g of vitamin E and 90 g of L-carnitine, and preparing 1,000packages of granules according to the method of Embodiment 1.

Embodiment 13

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract, 75 g of vitamin E and 100 g of L-carnitine, and preparing 1,000packages of granules according to the method of Embodiment 1.

Embodiment 14

Taking 105 g of silybin, 195 g of phospholipid, 300 g of Pu'er teaextract, 75 g of vitamin E and 100 g of L-carnitine, and preparing 1,000packages of granules according to the method of Embodiment 1.

Embodiment 15

Taking 120 g of silybin, 150 g of phospholipid, 360 g of Pu'er teaextract, 90 g of vitamin E and 120 g of L-carnitine, and preparing 1,000packages of granules according to the method of Embodiment 1.

Embodiment 16

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract, 75 g of vitamin E and 226.6 g of L-carnitine tartrate, andpreparing 1,000 packages of granules according to the method ofEmbodiment 1.

Embodiment 17

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract, 82.3 g of vitamin E and 100 g of L-carnitine, and preparing1,000 packages of granules according to the method of Embodiment 1.

Embodiment 18

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract, 92.4 g of vitamin E and 226.6 g of L-carnitine tartrate, andpreparing 1,000 packages of granules according to the method ofEmbodiment 1.

Embodiment 19

Taking the granules of Embodiment 8, adding 500 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 20

Taking the granules of Embodiment 9, adding 274 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 21

Taking the granules of Embodiment 10, adding 297 g of microcrystallinecellulose and 32 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 22

Taking the granules of Embodiment 13, adding 30 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 23

Taking the granules of Embodiment 14, adding 25 g of microcrystallinecellulose, mixing uniformly, encapsulated into capsules to obtain 1,000capsules.

Embodiment 24

Taking the composition of Embodiment 8, adding 400 g of lactose, 100 gof starch and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 25

Taking the composition of Embodiment 13, adding 20 g of lactose. 10 g ofsaponite and 64 g of low-substituted hydroxypropyl cellulose, mixinguniformly, encapsulated into capsules to obtain 1,000 capsules.

Embodiment 26

Taking the composition of Embodiment 13, adding 30 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,tablet pressing to obtain 1,000 tablets.

1. A pharmaceutical composition comprising: 8.75-60 parts by weight ofsilybin; 15-65 parts by weight of phospholipid; 25-200 parts by weightof Pu'er tea extract; 6.25-40 parts by weight of vitamin E; and 8.3-60parts by weight of L-carnitine.
 2. The pharmaceutical compositionaccording to claim 1 comprising: 25-40 parts by weight of silybin; 30-50parts by weight of phospholipid; 80-120 parts by weight of Pu'er teaextract; 10-30 parts by weight of vitamin E; and 25-40 parts by weightof L-carnitine.
 3. The pharmaceutical composition according to claim 2comprising: 35 parts by weight of silybin; 42 parts by weight ofphospholipid; 100 parts by weight of Pu'er tea extracts; 16.7 parts byweight of vitamin E; and 33.3 parts by weight of L-carnitine.
 4. Thepharmaceutical composition according to claim 1, wherein the vitamin Eis vitamin E acetate or vitamin E succinate.
 5. The pharmaceuticalcomposition according to claim 1, wherein the L-carnitine is L-carnitinetartrate.
 6. A pharmaceutical preparation comprising the pharmaceuticalcomposition according to claim 1, and pharmaceutically acceptablecarriers; wherein the pharmaceutically acceptable carriers are 0.1-99.9%of the total preparation by weight; the pharmaceutically acceptablecarriers are 40-70% of the total preparation by weight.
 7. Thepharmaceutical preparation according to claim 6, wherein thepharmaceutically acceptable carriers comprise one or more of mannitol,sorbitol, sorbic acid or sylvite, sodium metabisulfite, sodiumbisulfite, sodium thiosulfate, cysteine hydrochloride, mercaptoaceticacid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone,disodium EDTA, calcium disodium EDTA, the carbonate, acetate, phosphateof monovalence alkali metal or aqueous solution thereof, hydrochloricacid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodiumchloride, potassium chloride, sodium lactate, xylitol, maltose, glucose,fructose, dextran, glycine, starch, sucrose, lactose, mannitol, siliconderivative, cellulose and derivate thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, propylene glycol, ethanol, Tween 60-80, Span-80,beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters,agar, triethanolamine, basic amino acid, urea, allantoin, calciumcarbonate, calcium bicarbonate, surfactant, polyethylene glycol,cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talc,calcium stearate, magnesium stearate, or microcrystalline; preferably,wherein the carriers comprise one or more of microcrystalline cellulose,lactose, starch, sodium carboxymethylcellulose, low-substitutedhydroxypropyl cellulose, or talc.
 8. The pharmaceutical preparationaccording to claim 7, wherein the pharmaceutical preparation is any oneof tablet, sugar coated tablet, film coated tablet, enteric coatedtablet, capsule, hard capsule, soft capsule, oral liquid, oral agent,granule, pill, powder, paste, sublimed preparation, supensoid agent,solution, injection, suppository, ointment, emplastrum, creme, spray, orpatch; preferably, wherein the pharmaceutical preparation is a capsule,granule or tablet.
 9. A method of preparing the pharmaceuticalpreparation according to claim 6, comprising: (1) taking a prescriptionamount of raw materials for later use; (2) preparing a silybin complexliquid by weighing a prescription amount of silybin and phospholipin,and dissolving them in anhydrous ethanol, heating and refluxing toclarify the solution and continuing to heat for a time, thenconcentrating the clear solution under reduced pressure to aconcentrated volume, to obtain the silybin complex liquid for later use;(3) granulating by weighing a prescription amount of Pu'er tea extractas a base material, taking the silybin complex liquid prepared in step(2) as a feed liquid, and preparing granules by a fluidization spraymethod with a fluidized bed, and drying after the liquid complex is allsprayed in for later use; (4) achieving total blending by mixing thevitamin E, the L-carnitine and the granules prepared in step (3)uniformly to get the pharmaceutical composition; and (5) combining thepharmaceutical composition and the pharmaceutically acceptable carriersto make a conventional preparation.
 10. The method according to claim 9,wherein the heating time in step (2) is 0.5-1.5 hours; the concentratedvolume is 5%-20% of the original volume and the temperature ofconcentration under reduced pressure is 60-80° C.; the parameters of thefluidized bed in step (3) are that the temperature of the materials is40-65° C., and parameters such as fan frequency, the inlet airtemperature and the infusion frequency are adjusted to keep thematerials in a good fluidizing state during the granulation process; andwherein after granulation is completed, the granules are dried for 10-60minutes and the drying temperature is 55-65° C.
 11. Use of thepharmaceutical composition according to claim 1 in the preparation ofdrugs for treating non-alcoholic fatty liver diseases and/or reducingfat and losing weight and beautifying skin.
 12. A pharmaceuticalpreparation comprising the pharmaceutical composition according to claim2, and pharmaceutically acceptable carriers; wherein thepharmaceutically acceptable carriers are 0.1-99.9% of the totalpreparation by weight; the pharmaceutically acceptable carriers are40-70% of the total preparation by weight.
 13. The pharmaceuticalpreparation according to claim 12, wherein the pharmaceuticallyacceptable carriers comprise one or more of mannitol, sorbitol, sorbicacid or sylvite, sodium metabisulfite, sodium bisulfite, sodiumthiosulfate, cysteine hydrochloride, mercaptoacetic acid, methionine,vitamin A, vitamin C, vitamin E, vitamin D, azone, disodium EDTA,calcium disodium EDTA, the carbonate, acetate, phosphate of monovalencealkali metal or aqueous solution thereof, hydrochloric acid, aceticacid, sulfuric acid, phosphoric acid, amino acid, sodium chloride,potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose,dextran, glycine, starch, sucrose, lactose, mannitol, siliconderivative, cellulose and derivate thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, propylene glycol, ethanol, Tween 60-80, Span-80,beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters,agar, triethanolamine, basic amino acid, urea, allantoin, calciumcarbonate, calcium bicarbonate, surfactant, polyethylene glycol,cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talc,calcium stearate, magnesium stearate, or microcrystalline; preferably,wherein the carriers comprise one or more of microcrystalline cellulose,lactose, starch, sodium carboxymethylcellulose, low-substitutedhydroxypropyl cellulose, or talc.
 14. The pharmaceutical preparationaccording to claim 12, wherein the pharmaceutical preparation is any oneof tablet, sugar coated tablet, film coated tablet, enteric coatedtablet, capsule, hard capsule, soft capsule, oral liquid, oral agent,granule, pill, powder, paste, sublimed preparation, supensoid agent,solution, injection, suppository, ointment, emplastrum, creme, spray, orpatch; preferably, wherein the pharmaceutical preparation is a capsule,granule or tablet.
 15. A pharmaceutical preparation comprising thepharmaceutical composition according to claim 3, and pharmaceuticallyacceptable carriers; wherein the pharmaceutically acceptable carriersare 0.1-99.9% of the total preparation by weight; the pharmaceuticallyacceptable carriers are 40-70% of the total preparation by weight. 16.The pharmaceutical preparation according to claim 15, wherein thepharmaceutically acceptable carriers comprise one or more of mannitol,sorbitol, sorbic acid or sylvite, sodium metabisulfite, sodiumbisulfite, sodium thiosulfate, cysteine hydrochloride, mercaptoaceticacid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone,disodium EDTA, calcium disodium EDTA, the carbonate, acetate, phosphateof monovalence alkali metal or aqueous solution thereof, hydrochloricacid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodiumchloride, potassium chloride, sodium lactate, xylitol, maltose, glucose,fructose, dextran, glycine, starch, sucrose, lactose, mannitol, siliconderivative, cellulose and derivate thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, propylene glycol, ethanol, Tween 60-80, Span-80,beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters,agar, triethanolamine, basic amino acid, urea, allantoin, calciumcarbonate, calcium bicarbonate, surfactant, polyethylene glycol,cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talc,calcium stearate, magnesium stearate, or microcrystalline; preferably,wherein the carriers comprise one or more of microcrystalline cellulose,lactose, starch, sodium carboxymethylcellulose, low-substitutedhydroxypropyl cellulose, or talc.
 17. The pharmaceutical preparationaccording to claim 15, wherein the pharmaceutical preparation is any oneof tablet, sugar coated tablet, film coated tablet, enteric coatedtablet, capsule, hard capsule, soft capsule, oral liquid, oral agent,granule, pill, powder, paste, sublimed preparation, supensoid agent,solution, injection, suppository, ointment, emplastrum, creme, spray, orpatch; preferably, wherein the pharmaceutical preparation is a capsule,granule or tablet.
 18. A method of preparing the pharmaceuticalpreparation according to claim 12, comprising: (1) taking a prescriptionamount of raw materials for later use; (2) preparing a silybin complexliquid by weighing a prescription amount of silybin and phospholipin,and dissolving them in anhydrous ethanol, heating and refluxing toclarify the solution and continuing to heat for a time, thenconcentrating the clear solution under reduced pressure to aconcentrated volume, to obtain the silybin complex liquid for later use;(3) granulating by weighing a prescription amount of Pu'er tea extractas a base material, taking the silybin complex liquid prepared in step(2) as a feed liquid, and preparing granules by a fluidization spraymethod with a fluidized bed, and drying after the liquid complex is allsprayed in for later use; (4) achieving total blending by mixing thevitamin E, the L-carnitine and the granules prepared in step (3)uniformly to get the pharmaceutical composition; and (5) combining thepharmaceutical composition and the pharmaceutically acceptable carriersto make a conventional preparation.
 19. The method according to claim18, wherein the heating time in step (2) is 0.5-1.5 hours; theconcentrated volume is 5%-20% of the original volume and the temperatureof concentration under reduced pressure is 60-80° C.; the parameters ofthe fluidized bed in step (3) are that the temperature of the materialsis 40-65° C., and parameters such as fan frequency, the inlet airtemperature and the infusion frequency are adjusted to keep thematerials in a good fluidizing state during the granulation process; andwherein after granulation is completed, the granules are dried for 10-60minutes and the drying temperature is 55-65° C.
 20. Use of thepharmaceutical preparation according to claim 6 in the preparation ofdrugs for treating non-alcoholic fatty liver diseases and/or reducingfat and losing weight and beautifying skin.